RT-PCR扩增人抗HBsAg抗体Fab段基因

Amplification of Fab Fragments Genes of Human Antibodies against Hepatitis B Virus by Using RT-PCR

目的 采用逆转录 聚合酶链反应 (RT PCR)的方法从少量外周血中扩增抗HBsAg抗体重链Fd和κ轻链基因。方法 用RNAex试剂提取细胞总RNA ,以Olig (dT)引导合成cDNA第一链 ,再经PCR扩增的方法 ,以重链Fd和κ轻链基因简并引物 ,直接从人外周血混合淋巴细胞中扩增出抗HBsAg抗体重链Fd和κ轻链基因。 结果 扩增出分子量为 70 0bp的抗HBsAg抗体重链Fd和κ轻链基因。

Objective To obtain the anti hepatitis B virus (anti Hbs) antibody Fab genes by using RT PCR. Methods The total cell RNA extracted from peripheral lymphocytes of anti Hbs antibody positive volunteers was transcribed reversely into cDNA. The Fd fragments and light chain genes of human anti Hbs antibody were amplified from cDNA using polymerase chain reaction. Results Fd fragment gene and κchain gene of 700 bp was amplified. Conclusion Fab fragment genes may lay a solid foundation for further research into setting up recombinant adeno associated virus (r AAV).